RU2648990C1 - Lobaplatin crystals, methods of production and applications in pharmaceuticals - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: present invention relates to a novel crystal of a lobaplatin compound (crystalline form A). Crystal is characterised by two molecules of crystal water, which are in a crystal structure. Also disclosed are a method for preparation of crystal A, novel lobaplatin crystals, which are crystalline forms B and F, methods for preparation thereof, a pharmaceutical composition and use of said novel crystals for preparation of anticancer drugs.

EFFECT: in comparison with existing lobaplatin and lobaplatin trihydrate, lobaplatin crystals A, B and F have better stability and solubility and can be more effectively used in the treatment of cancer forms such as breast cancer, small cell lung cancer or chronic myeloid leukemia.

41 cl, 10 dwg, 15 tbl, 26 ex

Description

Technical field

The present invention relates to the field of pharmaceutical preparations, in particular, to new crystals of lobaplatin, methods for their preparation and their pharmaceutical applications, which are related to the field of medicines.

BACKGROUND OF THE INVENTION

Lobaplatin (D19466) is a third-generation platinum antitumor drug, after cisplatin and carboplatin, with the chemical name cis- [trans-1,2-cyclobutane bis (methylamine) -N, N '] - [(2S) -lactate- O1, O2] -platinum (II), the molecular formula C9H18N2O3Pt, a molecular weight of 397.34 and a chemical structural formula represented in the form of formula (1) as follows:

(1).

(one).

Lobaplatin refers to an alkylating agent (generically) with an alkanization function. Moreover, it has high antitumor activity, for example, it has a strong inhibitory effect on AH135 tumor in vitro, melanoma B16, colon tumor 115, mouse leukemia P338 in vivo, etc. Lobaplatin has the characteristics of strong anticancer activity, low toxicity, without cumulative toxicity and nephrotoxicity, in addition, low toxicity to bone marrow and without thrombocytopenia. Lobaplatin for injection, available for sale, is mainly used to treat breast cancer, small cell lung cancer and chronic myeloid leukemia.

The first prescription for this drug belongs to ASTA Pharmaceutical Co., Ltd. German company (ASTA Medica AG), for the first time the method for producing lobaplatin was described in the main patent EP 0324154. A method for producing lobaplatin trihydrate was disclosed in the subsequent patent EP 0611303, according to which the product is obtained by recrystallization of anhydrous lobaplatin with the formation of a crystalline product containing three molecules water. Patent EP 0611303 states that the lobaplatin obtained by the production method (from Example 1a) is characterized by a spreading property and thus lobaplatin is sticky and it is difficult to make preparations from it.

SUMMARY OF THE INVENTION

The technical problem solved by the present invention is that the currently existing anhydrous lobaplatin has several disadvantages regarding dispersion, great difficulties in its processing into drugs and lack of stability. The present invention provides new crystals of lobaplatin, which are most ideal for providing high bioavailability, good stability, high solubility, good fluidity, they are difficult to absorb moisture, therefore they are not sticky, and also have a good yield and purity.

A person skilled in the art knows that one and the same drug, having different crystalline forms, can have differences in bioavailability, as well as in stability, fluidity, compressibility, etc. Such physicochemical properties can have a certain effect on the use of medicines. Drug polymorphism has become a necessary and important part of the study of drugs, quality control and the testing process for pharmaceutical production. The study of drug polymorphism is advisable in relation to the biological activity selected among the new drug compounds, it is advisable in relation to improving bioavailability and enhancing the clinical therapeutic effect, it is advisable in relation to the selection and development of methods of drug administration and determination of multidisciplinary parameters for pharmaceutical preparations in order to thereby improve product quality.

Thanks to constant research and improvement, new crystalline forms of lobaplatin have been invented, as well as methods for their preparation and their use in pharmaceuticals.

More specifically, in order to solve the technical problems mentioned above, the present invention provides the following several types of technical solutions for new crystals of lobaplatin.

(1) Lobaplatin dihydrate

First, the present invention provides lobaplatin dihydrate (also called lobaplatin crystal A), methods for its preparation and its use in pharmaceuticals. Detailed descriptions are given below.

The present invention provides a crystal of a lobaplatin compound characterized by two molecules of crystalline water located in a crystal structure.

Preferably, the crystalline form of said lobaplatin compound of the present invention is crystalline Form A, and diffraction peaks are present on the PXRD pattern at 2θ values of about 11.04, 12.32, 12.61, 13.85, 15.14, 15 55, 16.68, 17.67, 17.86, 19.03, 20.06, 21.00, 22.68, 22.92, 23.76, 25.39, 25.58, 26.37 , 26.77, 27.00, 27.71, 28.13, 29.71, 31.42, 31.94, 32.89, 34.29, 34.60, 36.10, 36.93, 37 , 66, 40.78, 43.41, while the margin of error for the values of 2θ is 0.2.

Preferably, melting point T _like the specified crystal of the compound of lobaplatin is 220 ± 5 ° C.

Preferably, the crystalline form of said crystal of a lobaplatin compound is crystalline form A, which refers to an orthorhombic system with space group P2 ₁ 2 ₁ 2 ₁ , unit cell parameters a = 10.601 (2) Å, b = 14.020 (3), c = 9.759 ( 2) Å, α = β = γ = 90.0 °, unit cell volume V = 1450.5 (5) Å3, and the number of asymmetric units Z = 4 in the unit cell.

At the same time, the present invention further provides a method for producing the aforementioned lobaplatin compound, characterized in that it comprises the following steps:

adding lobaplatin trihydrate to the solvent to crystallize from the suspension to form a suspension mixture, stirring the mixture, precipitating the crystal, removing the solvent, and then drying to obtain a crystal.

Preferably, in the aforementioned method, after removing the solvent, the crystal is washed with ethyl ether before drying, wherein said drying is vacuum drying.

Preferably, in the aforementioned method, the weight ratio of said lobaplatin trihydrate to the volume of said crystallizing solvent is 1 (g): 15-30 (ml).

Preferably, in the aforementioned method, said crystallizing solvent is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane.

Preferably, in the above method, said suspension is obtained at room temperature, preferably, said suspension is obtained within 45-50 hours.

At the same time, the present invention provides a pharmaceutical composition, characterized in that the aforementioned lobaplatin compound is used as an active component.

Preferably, the amount of lobaplatin crystal in the minimum unit of said pharmaceutical composition is 5 mg, 10 mg or 50 mg.

Preferably, said pharmaceutical composition is in any clinically acceptable pharmaceutical dosage form.

Preferably, said dosage form is a lyophilized preparation for injection.

At the same time, the present invention further provides for the use of the aforementioned crystal of a lobaplatin compound or the aforementioned pharmaceutical composition for the manufacture of anti-cancer drugs.

At the same time, the present invention further provides the use of the aforementioned crystal of a lobaplatin compound or the aforementioned pharmaceutical composition for treating cancer, mainly for treating one of breast cancer, small cell lung cancer or chronic myeloid leukemia.

The present invention further provides for the use of the aforementioned crystalline form of lobaplatin for the preparation of pharmaceutical compositions and pharmaceutical preparations.

(2) Crystal B lobaplatin

Secondly, the present invention provides lobaplatin crystal B, methods for its preparation and its use in pharmaceuticals. Detailed descriptions are given below.

The present invention provides a crystal of a lobaplatin compound, characterized in that the crystalline form of the lobaplatin compound is crystalline Form B, and diffraction peaks are present on the PXRD pattern at 2θ values of about 8.25, 9.77, 11.70, 13.13, 15 , 28, 16.48, 17.22, 17.74, 19.01, 19.56, 22.28, 23.72, 24.04, 24.30, 25.62, 26.20, 28.57 , 30.22, 30.61, while the margin of error of the values of 2θ is 0.2.

Preferably, the melting point the specified crystal of the compound of lobaplatin according to the present invention is 230 ± 5 ° C.

The indicated melting point is measured by DSC with a heating rate of 10 ° C / min and evaluated by the maximum peak.

At the same time, the present invention further provides a method for producing a lobaplatin crystal with crystalline form B, comprising the following steps:

adding anhydrous methanol or anhydrous ethanol to lobaplatin trihydrate to obtain a mixture, stirring the mixture at room temperature to dissolve the solids, removing insoluble substances from the mixture, slowly evaporating the mixture, precipitating the crystal, isolating the crystal, and then drying the crystal to obtain a white powder, which represents a lobaplatin crystal B.

Preferably, in the above production method, the ratio of the mass of said lobaplatin trihydrate to the volume of said anhydrous methanol is 1 (g): 40-50 (ml); the ratio of the mass of the specified lobaplatin trihydrate to the volume of the indicated anhydrous ethanol is 1 (g): 80-90 (ml).

At the same time, the present invention further provides a method for producing a lobaplatin crystal with crystalline form B, comprising the following step b):

adding anhydrous methanol to lobaplatin dihydrate to obtain a mixture, stirring the mixture at room temperature to dissolve the solids, removing insoluble materials from the mixture, slowly evaporating the mixture, precipitating the crystal, isolating the crystal and drying the crystal to obtain a white powder, which is lobaplatin crystal B.

At the same time, the present invention further provides a method for producing a lobaplatin crystal with crystalline form B, which comprises the following step b):

adding an organic solvent to lobaplatin dihydrate to form a mixture in suspension, stirring the mixture at room temperature, precipitating a crystal, isolating a crystal and drying the crystal to give a white powder, which is lobaplatin crystal B.

Preferably, in the aforementioned production method, a method for producing said lobaplatin dihydrate comprises the following step a):

adding a crystallization solvent from the suspension to lobaplatin trihydrate to form a suspension, mixing the mixture, precipitating the crystal, washing with ethyl ether after removing the solvent, and then vacuum drying the crystal to obtain a crystal of lobaplatin dihydrate.

Preferably, in the aforementioned method, the ratio of the mass of said lobaplatin trihydrate to the volume of crystallizing solvent in step a) is 1 (g): 15-30 (ml).

Preferably, in the aforementioned method, said crystallization solvent in step a) is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane.

Preferably, in the aforementioned method, after isolating the crystal, the crystal is washed with ethyl ether before drying the crystal, where in step b) said drying is vacuum drying.

Preferably, in the aforementioned method, said suspension in step b) is obtained at room temperature, preferably, said suspension is obtained within 45-50 hours.

Preferably, in the aforementioned method, the ratio of the mass of lobaplatin dihydrate to the volume of anhydrous methanol in said step b) is 1 (g): 40-50 (ml).

Preferably, in the aforementioned method, the organic solvent in step b) is selected from n-hexane, acetone, ethyl acetate, nitromethane, acetonitrile, tetrahydrofuran, 2-butanone or dichloromethane, and the ratio of the mass of lobaplatin dihydrate to the volume of the organic solvent is 1 (g) : 15-30 (ml).

At the same time, the present invention provides a pharmaceutical composition, characterized in that the aforementioned lobaplatin crystal B is used as an active component.

Preferably, the amount of lobaplatin crystal in the minimum unit of said pharmaceutical composition is 5 mg, 10 mg or 50 mg.

Preferably, the form of said pharmaceutical composition is any clinically acceptable pharmaceutical dosage form.

Preferably, said dosage form is a lyophilized preparation for injection.

At the same time, the present invention further provides for the use of the aforementioned lobaplatin crystal or the aforementioned pharmaceutical composition for the manufacture of anticancer drugs.

At the same time, the present invention further provides for the use of the aforementioned crystal of a lobaplatin compound or the aforementioned pharmaceutical composition for treating cancer, in particular for treating one of breast cancer, small cell lung cancer or chronic myeloid leukemia.

(3) Crystal F Lobaplatin

Thirdly, the present invention provides a crystal F of lobaplatin, methods for its preparation and its use in pharmaceuticals. Detailed descriptions are given below.

The present invention provides a crystal of a lobaplatin compound, characterized in that the crystalline form of the compound is crystal F, and diffraction peaks are present on the PXRD pattern at 2θ values of about 8.21, 11.60, 12.99, 15.24, 16.44 , 17.11, 17.55, 18.42, 19.01, 19.20, 19.42, 21.81, 22.17, 22.42, 23.33, 23.85, 24.18, 24 , 40, 24.77, 25.46, 25.98, 26.13, 27.89, 28.42, 29.03, 30.32, 31.17, 31.94, 33.30, 36.20 , 37.62, 39.66, while the margin of error of the values of 2θ is 0.2.

Melting point T _like the specified crystal of the compound of lobaplatin is 229 ± 5 ° C.

Moreover, the indicated melting temperature is measured by DSC with a heating rate of 10 ° C./min and evaluated by the maximum peak.

At the same time, the present invention further provides a method for producing said lobaplatin crystal F, which is characterized in that it comprises the following step b):

adding methanol or ethanol to lobaplatin dihydrate to obtain a mixture, stirring the mixture at room temperature to dissolve the solids, filtering the mixture to remove insoluble substances, slowly adding the organic solvent to the mixture, precipitating the crystal, isolating the crystal, and then drying the crystal to obtain a white powder, which is lobaplatin crystal F.

Preferably, a method for producing said lobaplatin dihydrate comprises the following step a):

adding a suspension crystallization solvent to lobaplatin trihydrate to form a suspension, mixing the mixture, precipitating the crystal from the mixture, washing the crystal with ethyl ether after removing the solvent, and then vacuum drying the crystal to obtain a crystal of lobaplatin dihydrate.

Preferably, the weight ratio of said lobaplatin trihydrate to the volume of crystallization solvent in step a) is 1 (g): 15-30 (ml).

Preferably, said crystallization solvent in step a) is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane.

Preferably, in step b), after isolation of the crystal, the crystal is washed with ethyl ether before drying the crystal, wherein said drying is vacuum dehydration.

Preferably, said organic solvent in said step b) is selected from dimethyl ether of ethylene glycol, n-hexane, ethyl acetate, acetone, nitromethane, acetonitrile, tetrahydrofuran or methylene chloride.

Preferably, the weight ratio of said lobaplatin dihydrate to the volume of organic solvent in said step b) is 1 (g): 120-200 (ml).

Preferably, the weight ratio of said lobaplatin dihydrate to the volume of methanol is 1 (g): 40-50 (ml); the ratio of the mass of the specified lobaplatin dihydrate to the volume of ethanol is 1 (g): 80-90 (ml).

At the same time, the present invention provides a pharmaceutical composition, characterized in that the aforementioned lobaplatin crystal F is used as an active component.

Preferably, the amount of lobaplatin crystal in the minimum unit of said pharmaceutical composition is 5 mg, 10 mg or 50 mg.

Preferably, the form of said pharmaceutical composition is any clinically acceptable pharmaceutical dosage form.

Preferably, said dosage form is a lyophilized preparation for injection.

At the same time, the present invention further provides for the use of the aforementioned lobaplatin crystal or the aforementioned pharmaceutical composition for the manufacture of anticancer drugs.

At the same time, the present invention further provides for the use of the aforementioned lobaplatin crystal or the aforementioned pharmaceutical composition for treating cancer, in particular for treating one of breast cancer, small cell lung cancer or chronic myeloid leukemia.

The lobaplatin trihydrate starting material used in the present invention is prepared according to the method of the examples in patent EP 0611303.

The aforementioned lobaplatin crystal is used as an active ingredient in said pharmaceutical composition, and the amount of lobaplatin crystal in a minimum unit of said pharmaceutical composition is 5 mg, 10 mg or 50 mg. A new lobaplatin crystal can be prepared as a pharmaceutical composition with one or more pharmaceutically acceptable carriers or excipients. Furthermore, said pharmaceutical composition can be prepared in any of pharmaceutically acceptable dosage forms that are clinically suitable for the material, including non-gastrointestinal dosage forms, such as an injectable formulation, a transdermal formulation, respiratory dosage forms, formulations for mucosal administration through the cavity and other parts of the body, etc., mainly lyophilized powder for injection.

Pharmaceutical carriers or excipients may be selected from one or more of water for injection, mannitol, lactose, polyethylene glycols, polysorbate 80, propylene glycol, tartaric acid, citric acid, ascorbic acid, disodium edetate, sodium calcium edetate, sodium bisulfite, glucose, sodium chloride , soybean oil, soya lecithin, egg yolk phospholipids, distearyl phosphatidylethanolamine, dextran, glycine, glycerol.

A method of obtaining the above compositions and compositions is generally known to a person skilled in the art. Both the active form of the lobaplatin compounds of the present invention and the commercially available lobaplatin formulations are lobaplatin, and more specifically anhydrous lobaplatin, so that the lobaplatin compounds of the present invention are useful in treating all diseases in which commercially available lobaplatin-based products are used.

Lobaplatin, namely cis-platinum- [trans-1,2-cyclobutane-bis- (methylamine) -N, N '] - (2S) -lactate-O1, O2] -platinum (II), refers to an alkylating agent and cytotoxic drug, and is also known as a bioalkylating agent. Carbonium ions or other compounds having active electrophilic groups can be formed in vivo from lobaplatin and can be covalently linked to groups (such as amino, mercapto, hydroxyl, carboxyl group, phosphoric acid group, etc.) having a high content electrons in biological macromolecules (DNA, RNA, enzymes) of cells. Due to the loss of activity or rupture of DNA molecules as a result, the death of tumor cells can occur. This is why lobaplatin has strong antitumor activity. Pharmacokinetic studies show that after intravenous injection of lobaplatin all lobaplatin and the free lobaplatin that is formed have an antitumor effect in the serum, in other words, lobaplatin plays an effective role in this in the form of anhydrous lobaplatin, regardless of the state of the materials.

The above three types of new crystalline forms of lobaplatin of the present invention are designed to eliminate the disadvantages of amorphous lobaplatin, namely, easy diffusion, due to which it sticks, lack of stability, storage difficulties, etc. New crystalline forms of lobaplatin have characteristics such as high bioavailability , good stability, they are difficult to blur, etc. Moreover, it was unexpectedly discovered that new forms of lobaplatin have advantages over lobaplatin trihydrate, t Kie as high solubility, high yield and purity and better stability compared to lobaplatin trihydrate. Therefore, the creation of new crystalline forms is advisable in relation to the selection and development of methods of drug administration and determination of technological parameters for pharmaceutical preparations, so as to improve the quality of production of drugs. The new lobaplatin compounds of the present invention are very stable at normal temperatures, they are difficult to melt and therefore not sticky, they have good mobility, and compared with amorphous lobaplatin, they have the advantage of ease of use during storage, transportation, and preparation and processing.

Description of graphic materials

FIG. 1: X-ray diffraction pattern of lobaplatin dihydrate.

FIG. 2: Projection drawing of the molecular stereochemical structure of lobaplatin dihydrate.

FIG. 3: differential thermal analysis (DSC thermogram) of lobaplatin dihydrate.

FIG. 4: thermogravimetric analysis (TGA thermogram) of lobaplatin dihydrate.

FIG. 5: X-ray diffraction pattern of lobaplatin crystal B.

FIG. 6: differential thermal analysis (DSC thermogram) of lobaplatin crystal B.

7: thermogravimetric analysis (TGA thermogram) of lobaplatin crystal B.

FIG. 8: X-ray diffraction pattern of lobaplatin crystal F.

FIG. 9: differential thermal analysis (DSC thermogram) of lobaplatin crystal F.

Figure 10: thermogravimetric analysis (TGA thermogram) of lobaplatin crystal F.

Embodiments of the invention

(1) First Embodiment of the Present Invention

The present invention provides lobaplatin dihydrate characterized by a crystal morphology such that it is highly soluble, has a high yield and excellent stability. Specific descriptions are presented below.

The present invention provides crystalline Form A lobaplatin dihydrate and its identifying X-ray diffraction (PXRD) data are as follows.

The crystalline form A of lobaplatin dihydrate is analyzed using a Bruker D8 advance XRD X-ray diffractometer manufactured by Bruker. The measurement conditions are as follows: CuKa (40 kV, 40 mA), scan speed 2 ° / min (2θ values), scan range 3 ° -45 ° (2θ values). The crystalline form A of lobaplatin dihydrate is characterized by absorption peaks with the following characteristics shown in Table 1-a, and its X-ray diffraction pattern is shown in FIG. one.

Table 1-a. X-ray diffraction (PXRD) measurement result of crystalline form A of lobaplatin dihydrate

Peak Number 2θ diffraction angles (approximately) Interplanar distances, d (approximately) Relative intensity (approximately) one 11.04 8.01 66.9 2 12.32 7.18 86.3 3 12.61 7.01 51.3 four 13.85 6.39 22.2 5 15.14 5.85 one hundred 6 15,55 5.69 17.4 7 16.68 5.31 35 8 17.67 5.02 54.5 9 19.03 4.66 4.8 10 20.06 4.42 16.8 eleven 21.00 4.23 75.9 12 22.68 3.92 25,4 13 22.92 3.88 28,2 fourteen 23.76 3.74 2 fifteen 25.58 3.48 7.8 16 26.77 3.33 37,4 17 27.00 3.30 22.9 eighteen 27.71 3.22 8.3 19 28.13 3.17 19,2 twenty 29.71 3.00 8.4 21 31,42 2.84 33.5 22 31.94 2.80 25.1 23 32.89 2.72 8.9 23 34.60 2.59 17.3 25 36.93 2.43 10.3 26 37.66 2,39 7.3 27 40.78 2.21 12.3 28 43.41 2.08 9.8

The combination of lobaplatin with crystalline form A is a colorless and transparent columnar crystal, as determined by X-ray diffraction experiments. The combination of lobaplatin with crystalline form A refers to an orthorhombic syngony with space group P2 ₁ 2 ₁ 2 ₁ , unit cell parameters a = 10.601 (2), b = 14.020 (3), c = 9.759 (2) Å, α = β = γ = 90.0 °, unit cell volume V = 1450.5 (5) Å3, and the number of asymmetric units Z = 4 in the unit cell.

Diffraction intensity data are obtained using a Bruker SMART APEX-II diffractometer with the following parameters: CuKα radiation, graphite monochromator, diameter of an individual vessel f = 0.50 mm, distance between the crystal and CCD detector d = 60.3 mm, lamp voltage 40 kV , current of the electron tube 30 mA, scanning mode φ / ω scan. The number of all obtained diffraction points is 5844, of which the number of independent diffraction points is 2376, and the number of observed points is 2376 (| F | 2≥2σ | F | 2).

The crystal structure is analyzed by a direct method (Shelxs97), and all positions of 17 non-hydrogen atoms can be obtained. Structural parameters are refined, and atomic particles are determined using the least squares method. The positions of the hydrogen atoms are obtained by the method of geometric calculation and the difference Fourier method. And finally, the reliability factor R1 = 0.0569, wR2 = 0.1491 (w = 1 / σ | F | 2), S = 1,077. At the end, the stoichiometric formula C ₉ H ₁₈ N ₂ O ₃ Pt • 2H ₂ O was determined, with a calculated molecular weight of 433.36 and a calculated crystal density of 1.975 g / cm3. The resulting new lobaplatin crystal is lobaplatin dihydrate, determined by structural analysis, with the structure of the molecule represented by the following formula (2):

A projection drawing of the stereochemical structure of the molecule of lobaplatin dihydrate with crystalline form A is shown in figure 2.

The differential thermal analysis (DSC-TGA) of lobaplatin dihydrate with crystalline form A is carried out using differential thermal analyzers of the NETZSCH DSC 204 F1, NETZSCH TG 209 F1 models manufactured by NETZSCH. A DSC thermogram is shown in Figure 3, and a TGA thermogram is shown in Figure 4. The results show that the melting point T, measured by the DSC and estimated by the maximum peak, is the _like. was 220 ± 5 ° C, at a heating rate of 10 ° C / min. In particular, a wide endothermic peak of about 117 ° C is present in the DSC thermogram. This peak can be caused by the loss of two molecules of crystalline water, judging by the combination with data on the single crystal and TGA data. An exothermic peak at 220 ± 5 ° C is present on the DSC thermogram. This peak is the melting and decomposition peak, judging by the combination of the TGA data and the data on the melting temperature recorded in European patent EP 0611303. The TGA thermogram has a 9.49% weight loss before 150 ° C, which means that weight loss is caused by the loss of two molecules of crystalline water.

At the same time, the present invention provides a method for producing a new crystalline form A of lobaplatin, which is simple to obtain, easy to work with, and suitable for extended production, comprising the following steps:

weighing and placing lobaplatin trihydrate in a container, adding an organic solvent to the container to form a suspension mixture, stirring the mixture at room temperature for 45-50 hours, filtering the mixture, washing the product obtained by filtration with ethyl ether and then vacuum drying to obtain a white powder, which is lobaplatin dihydrate. The organic solvent is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane. The ratio of the mass of lobaplatin trihydrate to the volume of the organic solvent is 1 (g): 15-30 (ml).

(2) Second Embodiment of the Present Invention

The present invention provides a new lobaplatin crystal called crystalline Form B, which is highly soluble and has excellent stability.

On the one hand, the present invention provides lobaplatin crystal B, which is characterized by the morphology of crystal B. Exact descriptions of the obtained crystalline form B of lobaplatin according to the present invention are given below.

Lobaplatin crystalline Form B was analyzed using a Bruker D8 advance XRD X-ray diffractometer manufactured by Bruker. The measurement conditions are as follows: CuKa (40 kV, 40 mA), scan speed 2 ° / min (2θ values), scan range 3 ° -45 ° (2θ values). Crystalline form B of lobaplatin is characterized by absorption peaks with the following characteristics shown in Table 1-b, and its X-ray diffraction pattern is shown in FIG. 5.

Table 1-b. Measurement results of crystalline form B of lobaplatin by X-ray diffraction (PXRD)

Peak Number 2θ diffraction angles (approximately) Interplanar distance, d (approximately) Relative intensity (height, approximately,%) one 8.25 10.71 one hundred 2 9.77 9.04 3.6 3 11.70 7.55 4.7 four 13.13 6.74 16 5 15.28 5.80 13.7 6 16.48 5.37 8 7 17.22 5.15 8 8 17.74 4.99 20,2 9 19.01 4.66 27.4 10 19.56 4,53 14.1 eleven 22.28 3.99 7.4 12 23.72 3.75 6.1 13 24.04 3.70 8 fourteen 24.30 3.66 7.5 fifteen 25.62 3.47 3.6 16 26.20 3.40 5.9 17 28.57 3.12 3.3 eighteen 30.22 2.96 7.1 19 30.61 2.92 6.7

Differential thermal analysis (DSC-TGA) of the new crystalline form B of lobaplatin is carried out using differential thermal analyzers of the NETZSCH DSC 204 F1, NETZSCH TG 209 F1 models manufactured by NETZSCH. The DSC thermogram is shown in Figure 6, and the TGA thermogram is shown in Figure 7. The results show that the melting point T, measured by the DSC and evaluated by the maximum peak, _etc. was 230 ± 5 ° C, at a heating rate of 10 ° C / min. In particular, the DSC thermogram shows an exothermic peak at 230 ± 5 ° C, which is the melting and decomposition peak, judging by the combination of TGA data and data on the melting temperature recorded in European patent EP 0611303. There is no weight loss on the TGA thermogram before 150 ° C, this means that crystalline form B of lobaplatin is not a solvate.

At the same time, the present invention provides a method for producing a new crystalline Form B of lobaplatin, which is simple to obtain, easy to work with, and suitable for extended production.

In a preferred embodiment of the present invention, methods for producing said novel crystalline Form B of lobaplatin according to the present invention include the following steps:

a) obtaining lobaplatin dihydrate: weighing and placing lobaplatin trihydrate in a container, adding 15-30 ml of organic solvent per 1 g of lobaplatin trihydrate in a container to form a mixture in suspension, stirring the mixture at room temperature for 45-50 hours, filtering mixtures, washing the product obtained by filtration with ethyl ether, and then vacuum drying to obtain a white powder, which is lobaplatin dihydrate;

where the organic solvent is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane;

b) obtaining the desired crystal: weighing the lobaplatin dihydrate obtained in step a, placing the weighed lobaplatin dihydrate into the container, adding anhydrous methanol to the container, stirring at room temperature until the solids are dissolved, filtering to remove insoluble substances, slowly evaporating in a ventilated kitchen and isolating the crystal after the precipitation of the crystal, washing the crystal 2-3 times with ethyl ether, and then vacuum drying the crystal to obtain a white powder, which is It is a new crystalline form B lobaplatin.

The ratio of the mass of lobaplatin dihydrate to the volume of anhydrous methanol at the indicated stage b is 1 (g): 40-50 (ml).

In another preferred embodiment, the methods for producing said novel crystalline Form B of lobaplatin, preferably according to the present invention, may also be as follows:

a) obtaining lobaplatin dihydrate: weighing and placing lobaplatin trihydrate in a container, adding 15-30 ml of organic solvent in terms of 1 g of lobaplatin trihydrate in a container to form a mixture in suspension, stirring the mixture at room temperature for 45-50 hours, filtering the mixture, washing the product obtained by filtration with ethyl ether, and then vacuum drying to obtain a white powder, which is lobaplatin dihydrate;

where the organic solvent is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane;

b) obtaining the target crystal: weighing the lobaplatin dihydrate obtained in step a, placing the weighed lobaplatin dihydrate into the vessel, adding an organic solvent to the vessel to form a suspension mixture, stirring at room temperature for 45-50 hours, precipitating the crystal, filtering and isolating crystal, washing the crystal 2-3 times with ethyl ether, and then vacuum drying to obtain a white powder, which is a new crystalline form B of lobaplatin.

The specified organic solvent in step b is selected from n-hexane, acetone, ethyl acetate, nitromethane, acetonitrile, tetrahydrofuran, 2-butanone or dichloromethane. The ratio of the mass of lobaplatin dihydrate to the volume of the organic solvent is 1 (g): 15-30 (ml).

In another preferred embodiment, the methods for producing said new crystalline Form B of lobaplatin, preferably according to the present invention, may also be: adding anhydrous methanol or anhydrous ethanol to lobaplatin trihydrate, stirring at room temperature until the solids are dissolved, removing insoluble substances, slow evaporation , isolating the crystal after precipitation of the crystal, drying to obtain a white powder, which is a crystalline Form B lobaplatin. The ratio of the mass of lobaplatin trihydrate to the volume of anhydrous ethanol is 1 (g): 80-90 (ml).

(3) Third Embodiment of the Present Invention

The present invention provides a new crystal of lobaplatin, called crystalline form F, which is highly soluble and has excellent stability.

On the one hand, the present invention provides a crystal F of lobaplatin, which is characterized by the morphology of crystalline form F. Next, exact descriptions of the obtained crystalline form F of lobaplatin according to the present invention are given below.

Lobaplatin crystalline form F was analyzed using a Bruker D8 advance XRD X-ray diffractometer manufactured by Bruker. The measurement conditions are as follows: CuKa (40 kV, 40 mA), scan speed 2 ° / min (2θ values), scan range 3 ° -45 ° (2θ values). The crystalline form F of lobaplatin is characterized by absorption peaks with the following characteristics shown in Table 1-c, and its X-ray diffraction pattern is shown in FIG. 8.

Table 1-c. Measurement results of crystalline form F of lobaplatin by X-ray diffraction (PXRD)

Peak Number 2θ diffraction angles (approximately) Interplanar distance, d (approximately) Relative intensity (height, approximately,%) one 8.21 10.76 one hundred 2 11.60 7.62 0.8 3 12,99 6.81 3.3 four 15.24 5.81 5.2 5 16,44 5.39 8.9 6 17.11 5.18 2 7 17.55 5.05 2.2 8 18,42 4.81 0.9 9 19.01 4.67 6.3 10 19,20 4.62 2.2 eleven 19.42 4,57 2.1 12 21.81 4.07 0.7 13 22.17 4.01 1.7 fourteen 22.42 3.96 1.8 fifteen 23.33 3.81 0.8 16 23.85 3.73 3.2 17 24.18 3.68 1,2 eighteen 24.40 3.65 0.8 19 24.77 3,59 1.7 twenty 25.46 3,50 1,6 21 25.98 3.43 1.3 22 26.13 3.41 1,1 23 27.89 3.20 0.6 24 28.42 3.14 0.6 25 29.03 3.07 0.5 26 30.32 2.95 4.8 27 31.17 2.87 0.5 28 31.94 2.80 0.5 29th 33.30 2.69 1,1 thirty 36,20 2.48 1.8 31 37.62 2,39 1,2 32 39.66 2.27 0.8

Differential thermal analysis (DSC-TGA) of the new crystalline form F of lobaplatin is carried out using differential thermal analyzers of the NETZSCH DSC 204 F1, NETZSCH TG 209 F1 models manufactured by NETZSCH. The DSC thermogram is shown in Figure 9, and the TGA thermogram is shown in Figure 10. The results show that the melting point T, measured by the DSC and estimated by the maximum peak, is the _like. is 229 ± 5 ° C, at a heating rate of 10 ° C / min. In particular, the DSC thermogram has an exothermic peak at 229 ± 5 ° C, which is the melting and decomposition peak, judging by the combination of the TGA data and the data on the melting temperature recorded in European patent EP 0611303. A 1.97% loss is present on the TGA thermogram weight before 150 ° C, and when combined with DSC data, it can be concluded that weight loss is due to the remaining solvent.

At the same time, the present invention provides a method for producing a new crystalline form F of lobaplatin, which is simple to obtain, easy to work with, and suitable for extended production.

In a preferred embodiment of the present invention, methods for producing the new crystalline Form F of lobaplatin according to the present invention include the following steps:

a) obtaining lobaplatin dihydrate: weighing and placing lobaplatin trihydrate in a container, adding 15-30 ml of organic solvent in terms of 1 g of lobaplatin trihydrate in a container to form a mixture in suspension, stirring the mixture at room temperature for 45-50 hours, filtering the mixture, washing the product obtained by filtration with ethyl ether, and then vacuum drying to obtain a white powder, which is lobaplatin dihydrate;

where the organic solvent is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane;

b) obtaining the target crystal: weighing the lobaplatin dihydrate obtained in step a, placing the weighed lobaplatin dihydrate into the vessel, adding methanol or ethanol to the vessel to form a mixture, stirring the mixture at room temperature until the solids are dissolved, filtering to remove insoluble substances, slowly adding organic solvent to the filtered liquid, filtering and isolating the crystal after deposition of the crystal, washing the crystal with ethyl ether, and then vacuum drying Ivanov to give a white powder, which is a new crystalline form F lobaplatin.

The specified organic solvent in step b is selected from dimethyl ether of ethylene glycol, n-hexane, ethyl acetate, acetone, nitrotoluene, acetonitrile, tetrahydrofuran, dichloromethane. The ratio of the mass of lobaplatin dihydrate to the volume of the organic solvent is 1 (g): 120-200 (ml).

The mass ratio of lobaplatin dihydrate to the volume of methanol in the indicated step b is 1 (g): 40-50 (ml), and the ratio of the mass of lobaplatin dihydrate to the volume of methanol in the indicated step b is 1 (g): 80-90 (ml).

Examples

Methods for producing a lobaplatin crystal in forms A, B and F according to the present invention, as well as methods for screening and selecting various new crystalline forms, identifying and determining their properties are described in the following examples, in particular in the present invention.

Example 1. Screening and analysis of various crystalline forms

1.1. Screenings by means of a crystallization method by evaporation at room temperature

20 mg of a sample of lobaplatin trihydrate was taken in a 10 ml vial and 3 ml of anhydrous ethanol or anhydrous methanol was added to the vial to give a mixture. After complete dissolution of the lobaplatin trihydrate, the mixture was slowly evaporated at 25 ° C and dry solids were obtained. The resulting solids were analyzed using PXRD. The results are presented in table 2.

Table 2. Experiment results by crystallization by evaporation at room temperature

Rooms Solvents PXRD (probable crystalline form numbers) 1-1 Anhydrous ethanol B 1-2 Anhydrous methanol B

The results showed that, when compared, the crystalline form obtained using anhydrous methanol was the same as the crystalline form obtained using anhydrous ethanol, which was temporarily called crystalline form B.

1.2. Screenings by Suspension Crystallization Method

20 mg of a sample of lobaplatin trihydrate was taken in a 10 ml vial and 4 ml of the organic solvent mentioned in Table 3 was added to the vial to give a suspension. The resulting suspension was subjected to vibration for 5 hours at 25 ° C, and then the solvent was removed to obtain solids. After drying, the resulting solids were analyzed using PXRD. The results are presented in table 3.

Table 3. The results of the experiment on crystallization from suspension

The results showed that through crystallization from suspension, 9 kinds of crystalline forms were obtained, which were temporarily called crystalline forms A, B, C, D, E, G, H, I, L.

1.3. Screenings by Dissolution and Precipitation Crystallization Method

The solution was prepared by adding a 20 mg sample of lobaplatin trihydrate to 3 ml of anhydrous methanol or anhydrous ethanol. The following organic solvents were gradually added until solids precipitated, and then the supernatant was removed to obtain solids. After drying, solids were analyzed by PXRD. The results are presented in table 4.

Table 4. The results of the experiment on crystallization using solvent

The results showed that through crystallization with a solvent, 7 types of crystalline forms were obtained, which were temporarily called crystalline forms F, J, K, M, N, O, P.

1.4. Characterization of different crystalline forms

In addition to measuring PXRD for samples of crystalline forms of A-P, characterization using DCS and TGA can also be performed. The names, models and manufacturers of each of the tools used are presented in table 5.

Table 5. Models and manufacturers of each of the tools used to determine the characteristics of crystalline forms

The measurement results were as follows.

Crystalline Form A was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 11.04, 12.32, 12.61, 13.85, 15.14, 15.55, 16.68, 17.67, 17 86, 19.03, 20.06, 21.00.2 2.68, 22.92, 23.76, 25.39, 25.58, 26.37, 26.77, 27.00, 27, 71, 28.13, 29.71, 31.42, 31.94, 32.89, 34.29, 34.60, 36.10, 36.93, 37.66, 40.78, 43.41, the margin of error of the values of 2θ was 0.2.

Crystalline form A was characterized by the fact that an exothermic peak of about 220 ± 5 ° C was present in the DSC thermogram.

Crystalline Form B was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 8.25, 9.77, 11.70, 13.13, 15.28, 16.48, 17.22, 17.74, 19 , 01, 19.56, 22.28, 23.72, 24.04, 24.30, 25.62, 26.20, 28.57, 30.22, 30.61, while the error margin of 2θ values was 0.2.

Crystalline form B was characterized by the presence of an exothermic peak at about 230 ± 5 ° C in the DSC thermogram.

Crystalline Form C was characterized by the presence of diffraction peaks on a PXRD pattern at 2θ values of approximately 6.79, 8.07, 12.24, 12.61, 13.50, 16.50, 17.83, 18.32, 18 , 79, 20.09, 21.64, 22.27, 23.19, 24.73, 27.34, 28.35, 29.12, 31.92, while the margin of error of the 2θ values was 0.2.

Crystalline form C was characterized by the presence of an exothermic peak at about 228 ± 5 ° C in the DSC thermogram.

Crystalline form D was characterized by the fact that diffraction peaks were present on the PXRD pattern at 2θ values of approximately 6.76, 11.07, 12.35, 12.65, 13.88, 15.18, 15.56, 16.68, 17 70, 17.90, 20.08, 21.02, 22.70, 22.92, 25.41, 25.64, 26.41, 26.79, 27.02, 28.15, 31.44 , 31.96, 32.96, 34.34, 34.62, 36.93, 40.82, 43.46, while the margin of error of the 2θ values was 0.2.

Crystalline form D was characterized by the fact that an exothermic peak of about 218 ± 5 ° C was present on the DSC thermogram.

Crystalline form E was characterized by the fact that diffraction peaks were present on the PXRD pattern at 2θ values of approximately 6.61, 8.09, 12.38, 13.03, 15.40, 16.66, 17.47, 19.07, at the margin of error of the values of 2θ was 0.2.

Crystalline form E was characterized by the fact that an exothermic peak of about 214 ± 5 ° C was present in the DSC thermogram.

Crystalline Form F was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 8.21, 11.60, 12.99, 15.24, 16.44, 17.11, 17.55, 18.42, 19 01, 19.20, 19.42, 21.81, 22.17, 22.42, 23.33, 23.85, 24.18, 24.40, 24.77, 25.46, 25.98 , 26.13, 27.89, 28.42, 29.03, 30.32, 31.17, 31.94, 33.30, 36.20, 37.62, 39.66, while the margin of error 2θ was 0.2.

Crystalline form F was characterized by the fact that an exothermic peak of about 229 ± 5 ° C was present in the DSC thermogram.

Crystalline form G was characterized by the fact that diffraction peaks were present on the PXRD pattern at 2θ values of approximately 8.62, 10.82, 11.03, 12.26, 12.59, 13.82, 15.12, 15.57, 16 59, 17.43, 17.65, 18.48, 19.46, 20.11, 20.37, 21.01, 22.66, 22.86, 24.60, 25.40, 26.33 , 26.77, 27.00, 28.11, 29.79, 31.42, 31.94, 32.87, 34.25, 34.58, 36.06, 40.76, 42.75, 43 , 39, while the margin of error of the values of 2θ was 0.2.

Crystalline form H was characterized by the presence of diffraction peaks on a PXRD pattern at 2θ values of approximately 8.35, 8.53, 8.68, 12.97, 15.24, 17.41, 18.40, 19.13, 19 , 48, 20.37, 24.68, 25.41, 30.33, 31.66, 36.34, while the margin of error for the values of 2θ was 0.2.

Crystalline Form I was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 6.75, 8.39, 11.07, 11.59, 12.32, 12.63, 12.99, 15.20, 16 , 80, 17,07, 17,57,19,14,19,46, 21,00, 22,13, 22,84, 23,29, 23,77, 24,22, 25,82, 26,76 , 28.38, 30.34, 30.83, 31.90, 33.63, 36.32, 38.47, while the margin of error of the 2θ values was 0.2.

Crystalline form J was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 5.94, 8.35, 9.87, 13.05, 15.28, 16.66, 19.15, 22.22, 22 , 68, 25.09, 30.71, 33.56, while the margin of error for the values of 2θ was 0.2.

Crystalline form K was characterized by the fact that diffraction peaks were present on the PXRD pattern at 2θ values of approximately 8.29, 11.02, 12.31, 12.61, 13.84, 15.14, 15.53, 16.70, 17 66, 19.05, 20.06, 20.98, 22.68, 22.90, 25.60, 26.37, 26.77, 26.98, 27.68, 28.23, 29.75 , 31.40, 31.88, 32.90, 33.81, 34.29, 34.60, 36.10, 36.84, 37.64, 39.93, 40.76, 41.51, 42 , 36, 42.70, 43.39, while the margin of error for the values of 2θ was 0.2.

Crystalline form L was characterized by the fact that diffraction peaks were present on the PXRD pattern at 2θ values of approximately 6.71, 7.91, 10.75, 11.84, 14.06, 14.29, 15.85, 16.78, 17 , 29, 19.76, 20.20, 20.63, 21.08, 21.58, 21.89, 22.17, 23.87, 25.09, 26.83, 27.02, 28.73 , 29.18, 29.92, 30.56, 31.61, 33.95, 40.33, 41.33, while the margin of error of the 2θ values was 0.2.

Crystalline form M was characterized by the fact that diffraction peaks were present on the PXRD x-ray at 2θ values of approximately 8.05, 13.03, 15.20, 16.19, 17.47, 18.77, 19.32, 24.06, at the margin of error of the values of 2θ was 0.2.

Crystalline form N was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 7.94, 12.67, 14.83, 16.32, 17.16, 18.71, 21.83, 22.44, 24 , 10, 24.89, 27.97, 30.02, 30.48, while the margin of error for the values of 2θ was 0.2.

Crystalline form O was characterized by the fact that diffraction peaks were present on the PXRD x-ray at 2θ values of approximately 6.75, 8.15, 16.29, 18.95, 22.23, 24.52, 29.93, where the error margin was 2θ was 0.2.

Crystalline form P was characterized by the presence of diffraction peaks on the PXRD pattern at 2θ values of approximately 6.61, 8.17, 13.34, 16.52, 20.10, 24.97, 27.02, 33.99, 41 , 06, while the margin of error of the values of 2θ was 0.2.

1.5. Repeat and zoom experiments

For 100 mg of samples with A-P crystalline forms, extended and repeated tests were performed to verify the reproducibility of crystalline forms according to the aforementioned “1.2 Screenings by means of a crystallization method from suspension”. The results are presented in table 6 below.

Table 6. The results of repeated experiments and experiments on the increase in scale

The results showed that crystalline forms of A-F were stable. While some of the crystalline forms of G-P had difficulties with enlargement and some had a manifestation of the phenomenon of crystalline transformation, they were therefore unsuitable for further research.

Thus, screening of the starting materials was carried out by crystallization using methods of evaporation at normal temperatures, crystallization from suspension and crystallization by dissolution and precipitation, etc. X-ray diffraction patterns were analyzed after application of PXRD to determine the characteristics, and it was previously determined that lobaplatin may exist 16 kinds of crystalline forms of AP. After the verification procedure, by repeated experiments and scale-up experiments, it was determined that the crystalline forms of A-F were relatively stable with better reproducibility. Whereas with some of the other crystalline forms it was difficult to carry out scaled-up production with a lower yield, some concluded that these are unstable crystalline forms, since the phenomenon of crystalline transformation was observed. In this regard, crystalline forms of A-F were chosen further for a comprehensive study.

Example 2. Obtaining a new crystal of lobaplatin, called crystalline form A, as well as the properties of products from it and comparative analyzes

2.1. Obtaining lobaplatin dihydrate, called crystalline form A

Production Example 1

1 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of toluene was added to the container to obtain a suspension. The suspension was stirred at room temperature for 48 hours. Prior to precipitation of the crystals. The crystals were separated by filtration and washed 2-3 times with ethyl ether. Finally, after vacuum drying, 0.85 g of a white powder was obtained, which was lobaplatin dihydrate.

Production Example 2

1 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of ethyl ether was added to the container to obtain a suspension. The suspension was stirred at room temperature for 45 hours. Prior to precipitation of the crystals. The crystals were separated by filtration and washed 2-3 times with ethyl ether. Finally, after vacuum drying, 0.88 g of a white powder was obtained, which was lobaplatin dihydrate.

Production Example 3

1 g of lobaplatin trihydrate was weighed and placed in a container, and 20 ml of butyl acetate was added to the container to obtain a suspension. The suspension was stirred at room temperature for 50 hours. Prior to precipitation of the crystals. The crystals were separated by filtration and washed 2-3 times with ethyl ether. Finally, after vacuum drying, 0.83 g of a white powder was obtained, which was lobaplatin dihydrate.

Production Example 4

1 g of lobaplatin trihydrate was weighed and placed in a container, and 25 ml of 1,4-dioxane was added to the container to obtain a suspension. The suspension was stirred at room temperature for 48 hours until crystals precipitated. The crystals were separated by filtration and washed 2-3 times with ethyl ether. Finally, after vacuum drying, 0.90 g of a white powder was obtained, which was lobaplatin dihydrate.

Production Example 5

1 g of lobaplatin trihydrate was weighed and placed in a container, and 30 ml of n-heptane was added to the container to obtain a suspension. The suspension was stirred at room temperature for 46 hours. Before precipitation of the crystals. The crystals were separated by filtration and washed 2-3 times with ethyl ether. Finally, after vacuum drying, 0.87 g of a white powder was obtained, which was lobaplatin dihydrate.

Production Example 6

1 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of methyl tert-butyl ether was added to the container to give a suspension. The suspension was stirred at room temperature for 48 hours until crystals precipitated. The crystals were separated by filtration and washed 2-3 times with ethyl ether. Finally, after vacuum dehydration, 0.92 g of a white powder was obtained, which was lobaplatin dihydrate.

Samples obtained according to the aforementioned production examples were analyzed by XRD diffraction using method 1.4 in the above example 1. It was determined that all six samples had the same crystalline shape and characteristic peaks were as follows: diffraction peaks were present on the PXRD pattern at 2θ values of approximately 11, 04, 12.32, 12.61, 13.85, 15.14, 15.55, 16.68, 17.67, 17.86, 19.03, 20.06, 21.00, 22.68, 22.92, 23.76, 25.39, 25.58, 26.37, 26.77, 27.00, 27.71, 28.13, 29.71, 31.42, 31.94, 32, 89, 34.29, 34.60, 36.10, 36.93, 37.66, 40.78, 43.41, while the margin of error of the 2θ values was 0.2.

The results of elemental analyzes: C ₉ H ₁₈ N ₂ O ₃ Pt * 2H ₂ OM = 433.36

Calculated value (%): C 24.95 H 5.11 N 6.46 Pt 45.01

Measured value (%): C 24.94 H 5.08 N 6.41 Pt 45.07

This crystalline form was called crystalline form A.

2.2. Determination of the properties of products based on crystals of lobaplatin with crystalline form A and their comparative analyzes

1. Test samples

Lobaplatin dihydrate numbered 1-6 according to the present invention: lobaplatin dihydrate with crystalline form A was obtained using the method of Production Example 1-6, respectively.

Comparative sample 1: lobaplatin obtained using the method of example 1a, recorded in patent EP 0324154, and a specific production method are indicated below:

3.8 g (0.01 mol) of cis- [trans-1,2-butylbis (methylamine) -N, N '] dichloro-platinum (II) were suspended in 20 ml of water to give a mixture, and the mixture was heated up to 40 ° C. 3.39 g (0.02 mol) of silver nitrate was added to the mixture, and the mixture was stirred for 1.5 hours. The mixture was allowed to cool in the refrigerator to precipitate silver chloride. Precipitated silver chloride was then filtered and washed with 10 ml of water. The filtrate was passed through a column containing 100 ml of a basic ion exchanger and washed with 150 ml of water. Then, the filtrate was poured dropwise into 4.5 g (0.01 mol, 20% aqueous solution) of L-lactic acid. After stirring at room temperature for 3 days, the reaction mixture was concentrated, the residue was dissolved in methanol and stirred until it disappeared with additional activated carbon. Activated carbon was then filtered and ethyl ether was added to the filtrate. The solid obtained by rapid concentration was amorphous lobaplatin.

Comparative sample 2: lobaplatin trihydrate obtained according to the method recorded in the example of patent EP 0611303, and a specific method of preparation are indicated below:

3.8 g (0.01 mol) of cis- [trans-1,2-butyl bis (methylamine) -N, N '] dichloro-platinum (II) were suspended in 20 ml of water to give a mixture, and the mixture was heated to 40 ° C. 3.39 g (0.02 mol) of silver nitrate was added to the mixture, and the mixture was stirred for 1.5 hours. The mixture was allowed to cool in the refrigerator to precipitate silver chloride. The silver chloride precipitate was then filtered and washed with 10 ml of water. The filtrate was passed through a column containing 100 ml of a basic ion exchanger and washed with 150 ml of water. Then, the filtrate was poured dropwise into 4.5 g (0.01 mol, 20% aqueous solution) of L-lactic acid. After stirring at room temperature for 3 days, the reaction mixture was concentrated to approximately 20 ml and allowed to cool in the refrigerator overnight to precipitate a crystal. Then the crystal was filtered, and the filtrate was concentrated and left to cool in the refrigerator overnight again to precipitate the crystal. The crystal was filtered and the filtrate was collected. The crystals were combined and recrystallized from 20 ml of water / acetone (1/1, v / v), and the crystal obtained as a result was lobaplatin trihydrate.

2. Definition of morphology

Comparative sample 1: the obtained lobaplatin was amorphous;

Comparative Sample 2: Diffraction peaks were present on the PXRD pattern at 2θ values of about 6.71, 8.35, 12.89, 15.14, 16.74, 17.45, 19.01, 19.40, 22.07, 22.76, 23.16, 24.30, 25.21, 25.74, 27.08, 30.26, 30.79, while the margin of error of the 2θ values was less than 0.2. The melting point of this sample, component 210 ° C (decomposition), was described in patent EP 0611303.

Samples 1-6 with crystalline form A according to the present invention: diffraction peaks were present on the PXRD pattern at 2θ values of approximately 11.04, 12.32, 12.61, 13.85, 15.14, 15.55, 16.68, 17.67, 17.86, 19.03, 20.06, 21.00, 22.68, 22.92, 23.76, 25.39, 25.58, 26.37, 26.77, 27.00, 27.71, 28.13, 29.71, 31.42, 31.94, 32.89, 34.29, 34.60, 36.10, 36.93, 37.66, 40.78, 43.41, while the error margin of 2θ values was less than 0.2. A wide endothermic peak at about 117 ° C was present in the DSC thermogram, and this peak could be caused by the loss of two molecules of crystalline water, judging by the combination with the data on the single crystal and the TGA data. There was also an exothermic peak at 220 ± 5 ° C, which was the melting and decomposition peak, judging by the combination with the TGA data and the data on the melting temperature recorded in European patent EP 0611303. The TGA thermogram showed 9.49% weight loss before 150 ° C, which is caused by the loss of two molecules of crystalline water. All data showed that samples 1-6 had the same crystalline form, like lobaplatin dihydrate.

3. Solubility study

Lobaplatin trihydrate control solutions were prepared at concentrations of 60 μg / ml, 80 μg / ml, 200 μg / ml, 400 μg / ml and 800 μg / ml to make a standard curve using the HPLC method. The resulting standard curve was Y = 4.8641X + 20.5794, R = 0.9998. Saturated aqueous solutions (suspensions) of sample 6 of lobaplatin dihydrate and comparative sample 2, respectively, were prepared. The solutions were vibrated for 6 hours in a shaker at 25 ° C, followed by filtration and diluted to the appropriate multiples for analysis using HPLC. The solubility results are presented in table 7 below.

Table 7. The results of the study of solubility

Crystalline form Sample 6 Comparative Sample 2 Solubility (mg / ml) 16,7307 10,3271

The results showed that the solubility of lobaplatin dihydrate obtained according to the present invention was better than that of lobaplatin trihydrate.

4. The study of the quality of new crystalline forms of lobaplatin by comparison

20 mg of samples 1-6 of lobaplatin dihydrate and comparative samples 1-2 were weighed, respectively. Values of moisture content, impurities, content of active ingredients and product yields were considered as indicators for studying the quality and yields of products. The results are presented in table 8 below.

Table 8. The study of the quality of lobaplatin dihydrate with crystalline form A by comparison

The above results showed that, compared with anhydrous lobaplatin and lobaplatin trihydrate, the new lobaplatin A crystal obtained according to the present invention has characteristics of high contents, low impurity contents and good yields.

Note 1. The method of determining the values of the content: measured using high performance liquid chromatography; chromatographic conditions were as follows: silica gel grafted with octadecyl silane as a filler, the ratio of potassium dihydrogen phosphate solution to acetonitrile 92: 8 as the mobile phase, the detection wavelength of 210 nm, the column temperature was 40 ° C, the number of theoretical plates calculated from the lobaplatin peaks, must be at least 1000 and the degree of separation of lobaplatin peaks and impurity peaks must meet the requirements. Preparation of a control product solution: 10 mg of a product based on lobaplatin trihydrate for control was accurately weighed and fixed in a 50 ml volumetric flask, diluted to the mark with water and shaken well. Preparation of test solutions: 20 mg of the sample was precisely weighed in the indicated order and fixed in a 100 ml volumetric flask, diluted to the mark with water and shaken well. Measurements and results: exactly 10 μl of the solutions of the product for control and the solutions of the sample were measured, respectively, introduced into a liquid chromatograph, and chromatograms were recorded. The content values calculated taking into account peak areas using the external standard method were calculated relative to anhydrous lobaplatin with a standard range of 97.0% -102%.

Note 2. A method for checking for the presence of impurities: lobaplatin, 1,2-diaminomethylcyclobutane (CBMA), lactic acid and other known and unknown impurities were determined using TLC. Developer: ethanol: chloroform: 25% aqueous ammonia: water = 53: 39: 15: 1.5 (volume ratio), plates for TLC: silica gel 60 F25410 × 10 plates for TLC. Plates were placed in iodine vapors for staining with 0.3% ninhydrin reagent and chromogenic reagent nitrosodimethylaniline after development. Checked for the presence of CBMA and unknown impurities.

Note 3. The method of determining the values of moisture content: used the Karl Fischer method. The theoretical moisture content for lobaplatin dihydrate was 8.77%, and the theoretical moisture content for lobaplatin trihydrate was 11.96%.

5. Stability study

Sample 6 obtained in the examples of the present invention and the resulting comparative sample 2 were left under the following conditions: furnace temperature of 60 ° C, relative humidity of approximately 95%, and a light stability test tank of approximately 4,500 lux (conventional thermostat with lighting function) . After 5 days and 10 days, the samples were removed for PXRD tests and HPLC assays in order to examine the stability of the samples under conditions of high temperature, high humidity and light. The results are presented in table 9.

Table 9. The results of the stability assessment

The above experimental results showed that the lobaplatin according to the present invention was characterized by a new crystalline form, which had a higher solubility than the solubility of lobaplatin trihydrate, while the yields and purity were ideal. The new crystalline form of lobaplatin had good stability and did not show the phenomenon of crystalline transformation according to the results of the study at high temperature, high humidity and lighting. The content of active ingredients was higher than that of lobaplatin trihydrate, and had no visible changes according to the HPLC results, meaning that the new crystalline form of lobaplatin according to the present invention was more stable than the crystalline form of lobaplatin, it was difficult to dissolve, therefore it was not sticky, and had good fluidity.

Example 3. Obtaining a new crystal of lobaplatin, called crystalline form B, as well as the properties of products from it and comparative analyzes

3.1. Obtaining a new crystal of lobaplatin called crystalline form B

Production Example 1

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 30 ml of toluene was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.73 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the desired crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 40 ml of anhydrous methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, slowly evaporated in a fume hood until a crystal precipitated, and then the crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.74 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 2

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether and then, after vacuum drying, 1.84 g of a white powder, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 50 ml of anhydrous methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, slowly evaporated in a fume hood until a crystal precipitated, and then the crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.76 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 3

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 20 ml of butyl acetate was added to the container to obtain a suspension. The suspension was stirred for 50 hours at room temperature, filtered, and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.68 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 20 ml of n-hexane was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.75 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 4

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 25 ml of 1,4-dioxane was added to the container to obtain a suspension. The suspension was stirred for 45 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.76 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 15 ml of acetone was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.78 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 5

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 30 ml of n-heptane was added to the container to obtain a suspension. The suspension was stirred for 50 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.75 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 18 ml of ethyl acetate was added to the container to obtain a suspension. The suspension was stirred for 50 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.75 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 6

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of ethyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.78 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the desired crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 25 ml of nitromethane was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.77 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 7

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 18 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered, and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.82 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 30 ml of tetrahydrofuran was added to the container to obtain a suspension. The suspension was stirred for 46 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.79 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 8

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 25 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 46 hours at room temperature, filtered, and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.83 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 15 ml of dichloromethane was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.76 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 9

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 30 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered, and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.85 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 25 ml of acetonitrile was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.78 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 10

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.84 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 20 ml of 2-butanonas was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed 2-3 times with ethyl ether, and after vacuum drying, 0.73 g of a white powder was obtained, which was a new crystalline Form B of lobaplatin.

Production Example 11

1 g of lobaplatin trihydrate was weighed and placed in a container, and 40 ml of anhydrous methanol was added to the vessel to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then the mixture was slowly evaporated to precipitate a crystal. The crystal was filtered and isolated. After drying, 0.68 g of a white powder was obtained, which was crystalline Form B of lobaplatin.

Production Example 12

1 g of lobaplatin trihydrate was weighed and placed in a container, and 85 ml of anhydrous ethanol was added to the vessel to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then the mixture was slowly evaporated to precipitate a crystal. The crystal was filtered and isolated. After drying, 0.70 g of a white powder was obtained, which was crystalline Form B of lobaplatin.

Samples obtained according to stage a) in the above production examples 1-10 were analyzed by XRD diffraction by method 1.4 in the above example 1. It was determined that all 10 samples had the same crystalline form, which was crystalline form A. And detailed specific data were the same as in the previous first part of the embodiment, which is the “first embodiment of the present invention”, and as in the previous example 2.

At the same time, the samples obtained by stage b) in the above production examples 1-10 and examples 11-12 were analyzed using XRD diffraction by method 1.4 in the above example 1. It was determined that all 12 samples had the same crystalline shape, and characteristic the peaks were as follows: diffraction peaks were present on the PXRD pattern at 2θ values of approximately 8.25, 9.77, 11.70, 13.13, 15.28, 16.48, 17.22, 17.74, 19.01, 19.56, 22.28, 23.72, 24.04, 24.30, 25.62, 26.20, 28.57, 30.22, 30.61, while the margin of error of the 2θ values was 0.2 . This crystalline form was called crystalline form B.

3.2. Determination of the properties of products with crystalline form B and their comparative analyzes

1. Test samples

Samples 1-12: compounds of lobaplatin with crystalline form B were obtained by the method of Production Examples 1-12 in Example 3 of the present invention, respectively.

Comparative sample 1: was the same as the previous sample from comparative sample 1 in part 2.2 of example 2. Lobaplatin obtained by the method of example 1a in patent EP 0324154, and the specific preparation methods were the same as in the previous comparative sample 1 in part 2.2 of example 2.

Comparative sample 2: was the same as the previous sample from comparative sample 2 in part 2.2 of example 2. Lobaplatin trihydrate was obtained by the method of example in patent EP 0611303, and the specific production methods were the same as in the case of the previous comparative sample 2 in part 2.2 of example 2.

2. Definition of morphology

The results of the determination were the same as that of comparative sample 1 and comparative sample 2 in part 2.2 of example 2, in particular, the results are listed below:

Comparative sample 1: the obtained lobaplatin was amorphous;

Comparative Sample 2: Diffraction peaks were present on the PXRD pattern at 2θ values of about 6.71, 8.35, 12.89, 15.14, 16.74, 17.45, 19.01, 19.40, 22.07, 22.76, 23.16, 24.30, 25.21, 25.74, 27.08, 30.26, 30.79, while the margin of error of the 2θ values was less than 0.2. The melting point of this sample, component 210 ° C (decomposition), was described in patent EP 0611303.

Samples 1-12 according to the present invention: diffraction peaks were present on the PXRD pattern at 2θ values of approximately 8.25, 9.77, 11.70, 13.13, 15.28, 16.48, 17.22, 17.74, 19.01, 19.56, 22.28, 23.72, 24.04, 24.30, 25.62, 26.20, 28.57, 30.22, 30.61, while the error margin is 2θ was less than 0.2. There was also an exothermic peak at 230 ± 5 ° C, which was the melting and decomposition peak, judging by the combination with the TGA data and the data on the melting temperature recorded in European patent EP 0611303. All data showed that samples 1-12 had the same crystalline shape which was the new crystalline form B of lobaplatin.

3. Solubility study

Lobaplatin trihydrate control solutions were prepared at concentrations of 60 μg / ml, 80 μg / ml, 200 μg / ml, 400 μg / ml and 800 μg / ml to make a standard curve using the HPLC method. The resulting standard curve was Y = 4.8641X + 20.5794, R = 0.9998. Saturated aqueous solutions (suspensions) of sample 1 were prepared with a new crystalline form B of lobaplatin and comparative sample 2, respectively. The solutions were vibrated in a shaker at 25 ° C for 6 hours, followed by filtration and diluted to appropriate multiplicities for analysis by HPLC. The solubility results are presented in table 10 below.

Table 10. The results of the study of solubility

Crystalline form Sample 1 Comparative Sample 1 Solubility (mg / ml) 17,7341 10,3271

The results showed that the solubility of the lobaplatin compound obtained according to the present invention was better than that of lobaplatin trihydrate.

4. The study of the quality of new crystalline forms of lobaplatin by comparison

50 mg of samples 1-12 of lobaplatin compounds and comparative samples 1-2 were weighed, respectively. Values of moisture content, impurities, content of active ingredients and product yields were considered as indicators for studying the quality and yields of products. The theoretical moisture content for lobaplatin trihydrate was 11.96%. The results are presented in table 11 below.

Table 11. The study of the quality of the new crystalline form B of lobaplatin by comparison

The above results showed that, compared to anhydrous lobaplatin and lobaplatin trihydrate, the new crystalline form B of lobaplatin obtained according to the present invention has characteristics of high content, low impurity content and good yields.

Note 1. The method for determining the content values: was the same as in the previous part 2.2 of example 2.

Note 2. The method of checking for the presence of impurities: was the same as in the previous part 2.2 of example 2.

Note 3. The method of determining the values of moisture content: was the same as in the previous part 2.2 of example 2.

5. Stability study

Sample 1, obtained in example 3 of the present invention, and the obtained comparative sample 2 were left under the following conditions: furnace temperature of 60 ° C, relative humidity of approximately 95%, and light stability test tank for lighting of approximately 4,500 lux (conventional thermostat with lighting function) . After 5 days and 10 days, the samples were removed for PXRD tests and HPLC assays in order to examine the stability of the samples under conditions of high temperature, high humidity and light. The results are presented in table 12.

Table 12. The results of the stability assessment

The above experimental results showed that the new crystalline form B of lobaplatin according to the present invention had a higher solubility than the solubility of lobaplatin trihydrate, while the yields and purity were ideal. The new crystalline form B of lobaplatin had good stability and did not exhibit the phenomenon of crystalline transformation according to the results of the study at high temperature, high humidity and lighting. The content of active ingredients was higher than that of lobaplatin trihydrate, and had no visible changes according to the HPLC results, meaning that the new crystalline form of lobaplatin according to the present invention was more stable than the crystalline form of lobaplatin, it was difficult to dissolve, therefore it was not sticky, and had good fluidity.

Example 4. Obtaining a new crystal of lobaplatin, called crystalline form A, as well as the properties of products from it and comparative analyzes

4.1. Obtaining a new crystal of lobaplatin called crystalline form F

Production Example 1

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 30 ml of toluene was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.73 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 40 ml of methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 120 ml of ethylene glycol dimethyl ether was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.71 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 2

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.84 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 50 ml of methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 150 ml of n-hexane was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.68 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 3

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 20 ml of butyl acetate was added to the container to obtain a suspension. The suspension was stirred for 50 hours at room temperature, filtered, and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.68 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 80 ml of ethanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 200 ml of ethyl acetate was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.70 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 4

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 25 ml of 1,4-dioxane was added to the container to obtain a suspension. The suspension was stirred for 45 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.76 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 90 ml of ethanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 180 ml of acetone was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.72 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 5

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 30 ml of n-heptane was added to the container to obtain a suspension. The suspension was stirred for 50 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.75 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 45 ml of methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 160 ml of nitromethane was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.69 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 6

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of ethyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.78 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 40 ml of methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 150 ml of acetonitrile was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.73 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 7

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 15 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 48 hours at room temperature, filtered and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.84 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 85 ml of ethanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 180 ml of tetrahydrofuran was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether, and after vacuum drying, 0.67 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Production Example 8

a) Preparation of lobaplatin dihydrate: 2 g of lobaplatin trihydrate was weighed and placed in a container, and 20 ml of methyl tert-butyl ether was added to the container to obtain a suspension. The suspension was stirred for 46 hours at room temperature, filtered, and the product obtained by filtration was washed with ethyl ether, and then, after vacuum drying, 1.80 g of a white powder was obtained, which was lobaplatin dihydrate.

b) Preparation of the target crystal: 1 g of lobaplatin dihydrate from stage a was weighed and placed in a container, and 40 ml of methanol was added to the container to obtain a mixture. The mixture was stirred at room temperature until the solids dissolved. The mixture was filtered to remove insoluble matter, and then 150 ml of dichloromethane was slowly added to the mixture to precipitate a crystal. The crystal was filtered and isolated. Then, the crystal was washed with ethyl ether and, after vacuum drying, 0.66 g of a white powder was obtained, which was a new crystalline form F of lobaplatin.

Samples obtained according to stage a) in the above production examples 1-8 were analyzed by XRD diffraction by method 1.4 in the above example 1. It was determined that all 8 samples had the same crystalline form, which was crystalline form F. And detailed specific data were the same as in the previous first part of the embodiment, which is the "first embodiment of the present invention", and as in the previous example 2.

At the same time, the samples obtained in the above production examples 1-8 by the method 1.4 of Example 1 were analyzed using XRD diffraction. It was determined that all 8 samples had the same crystalline shape, and the characteristic peaks were as follows: diffraction peaks were present on the PXRD pattern at 2θ values of approximately 8.21, 11.60, 12.99, 15.24, 16.44, 17.11 , 17.55, 18.42, 19.01, 19.20, 19.42, 21.81, 22.17, 22.42, 23.33, 23.85, 24.18, 24.40, 24 , 77, 25.46, 25.98, 26.13, 27.89, 28.42, 29.03, 30.32, 31.17, 31.94, 33.30, 36.20, 37.62 , 39.66, while the margin of error of the values of 2θ was 0.2. This crystalline form was called crystalline form F.

4.2. Determination of the properties of products with crystalline form F and their comparative analyzes

1. Test samples

Samples 1-8: obtained by the method of Production Examples 1-8 in the present invention, respectively.

Comparative sample 1: was the same as the previous sample from comparative sample 1 in part 2.2 of example 2. Lobaplatin obtained according to the method of example 1a in patent EP 0324154, and the specific preparation methods were the same as the previous comparative sample 1 in part 2.2 of example 2.

Comparative sample 2: was the same as the previous sample from comparative samples 2 in part 2.2 of example 2. Lobaplatin was obtained according to the method of example in patent EP 0611303, and the specific preparation methods were the same as in the case of the previous comparative sample 2 in part 2.2 example 2.

2. Definition of morphology

The results of the determination were the same as that of comparative sample 1 and comparative sample 2 in part 2.2 of example 2, in particular, the results are listed below.

Comparative sample 1: the obtained lobaplatin was amorphous;

Comparative Sample 2: Diffraction peaks were present on the PXRD pattern at 2θ values of about 6.71, 8.35, 12.89, 15.14, 16.74, 17.45, 19.01, 19.40, 22.07, 22.76, 23.16, 24.30, 25.21, 25.74, 27.08, 30.26, 30.79, while the margin of error of the 2θ values was less than 0.2. The melting point of this sample, component 210 ° C (decomposition), was described in patent EP 0611303.

Samples 1-8 according to the present invention: diffraction peaks were present on the PXRD pattern at 2θ values of approximately 8.21, 11.60, 12.99, 15.24, 16.44, 17.11, 17.55, 18.42, 19.01, 19.20, 19.42, 21.81, 22.17, 22.42, 23.33, 23.85, 24.18, 24.40, 24.77, 25.46, 25, 98, 26.13, 27.89, 28.42, 29.03, 30.32, 31.17, 31.94, 33.30, 36.20, 37.62, 39.66, while the margin of error 2θ values was less than 0.2. The DSC thermogram showed an exothermic peak at about 229 ° C, which was the melting and decomposition peak, judging by the combination with the TGA data and the data on the melting temperature recorded in European patent EP 0611303. All data showed that samples 1-8 had the same crystalline form which was the new crystalline form F of lobaplatin.

3. Solubility study

Lobaplatin trihydrate control solutions were prepared at concentrations of 60 μg / ml, 80 μg / ml, 200 μg / ml, 400 μg / ml and 800 μg / ml to make a standard curve using the HPLC method. The resulting standard curve was Y = 4.8641X + 20.5794, R = 0.9998. Prepared saturated aqueous solutions (suspensions) of sample 6 of a new crystal of lobaplatin and comparative sample 2, respectively. The solutions were vibrated in a shaker at 25 ° C for 6 hours, followed by filtration and diluted to appropriate multiplicities for analysis by HPLC. The solubility results are presented in table 13 below.

Table 13. Solubility Results

Crystalline form Sample 6 Comparative Sample 1 Solubility (mg / ml) 21,4957 10,3271

The results showed that the solubility of the lobaplatin compound obtained according to the present invention was better than that of lobaplatin trihydrate.

4. The study of the quality of new crystalline forms of lobaplatin by comparison

20 mg of samples 1-8 of lobaplatin dihydrate and comparative sample 1-2 were weighed, respectively. Values of moisture content, impurities, content of active ingredients and product yields were considered as indicators for studying the quality and yields of products. The results are presented in table 14 below.

Table 14. The study of the quality of a new crystal of lobaplatin by comparison

The above results showed that, compared to anhydrous lobaplatin and lobaplatin trihydrate, the new crystalline form F of lobaplatin obtained according to the present invention is characterized by high contents, low impurity contents and good yields.

Note 1. The method for determining the content values: was the same as in the previous part 2.2 of example 2.

Note 2. The method of checking for the presence of impurities: was the same as in the previous part 2.2 of example 2.

Note 3. The method of determining the values of moisture content: was the same as in the previous part 2.2 of example 2.

5. Stability study

Sample 6, obtained in the example of the present invention, and the obtained comparative sample 2 were left under the following conditions: furnace temperature of 60 ° C, relative humidity of approximately 95%, and light stability test tank for lighting of approximately 4,500 lux (conventional thermostat with lighting function) . After 5 days and 10 days, the samples were removed for PXRD tests and HPLC assays in order to examine the stability of the samples under conditions of high temperature, high humidity and light. The results are presented in table 15.

Table 15. The results of the stability assessment

The above experimental results showed that the new crystalline form F of lobaplatin according to the present invention had a higher solubility than the solubility of lobaplatin trihydrate, while the yields and purity were ideal. The new crystalline form of lobaplatin F had good stability and did not exhibit the phenomenon of crystalline transformation according to the results of the study at high temperature, high humidity and light. The content of active ingredients was higher than that of lobaplatin trihydrate, and had no visible changes according to HPLC results, meaning that the new lobaplatin crystal was stable.

Claims (49)

1. A crystal of lobaplatin compound, characterized by two molecules of crystalline water located in the crystal structure. 2. The crystal of the lobaplatin compound of claim 1, wherein the crystalline form of said lobaplatin compound is crystalline form A and diffraction peaks are present on the PXRD pattern at 2θ values of about 11.04, 12.32, 12.61, 13.85, 15.14, 15.55, 16.68, 17.67, 17.86, 19.03, 20.06, 21.00, 22.68, 22.92, 23.76, 25.39, 25, 58, 26.37, 26.77, 27.00, 27.71, 28.13, 29.71, 31.42, 31.94, 32.89, 34.29, 34.60, 36.10, 36.93, 37.66, 40.78, 43.41, while the margin of error for the values of 2θ is 0.2. 3. The crystal of lobaplatin compound according to claim 2, where the melting point is T, _etc. the specified crystal of the compound of lobaplatin is 220 ± 5 ° C. 4. The lobaplatin compound crystal according to claim 1, wherein the crystalline form of said lobaplatin compound crystal is crystalline form A, which refers to an orthorhombic system with space group P2 ₁ 2 ₁ 2 ₁ , unit cell parameters a = 10,601 (2) Å, b = 14.020 (3), c = 9.759 (2) Å, α = β = γ = 90.0 °, unit cell volume V = 1450.5 (5) Å3, and the number of asymmetric units Z = 4 in the unit cell. 5. A method of obtaining the specified compound lobaplatin according to any one of paragraphs. 1-4, characterized in that the method includes the following stages: adding lobaplatin trihydrate to the solvent to crystallize from the suspension to form a suspension mixture, stirring the mixture, precipitating the crystal, removing the solvent, and then drying to obtain a crystal. 6. The method according to p. 5, where after removing the solvent, the crystal is washed with ethyl ether before drying, and the specified drying is a vacuum drying. 7. The method according to p. 5 or 6, where the mass ratio of the specified lobaplatin trihydrate to the volume of the crystallization solvent is 1 (g): 15-30 (ml). 8. The method according to any one of paragraphs. 5, 6, wherein said crystallization solvent is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane. 9. The method according to any one of paragraphs. 5, 6, where the specified suspension is obtained at room temperature, preferably the specified suspension is obtained within 45-50 hours 10. A crystal of lobaplatin compound, characterized in that the crystalline form of said lobaplatin compound is crystalline form B and diffraction peaks are present on the PXRD pattern at 2θ values of approximately 8.25, 9.77, 11.70, 13.13, 15 , 28, 16.48, 17.22, 17.74, 19.01, 19.56, 22.28, 23.72, 24.04, 24.30, 25.62, 26.20, 28.57 , 30.22, 30.61, while the margin of error of the values of 2θ is 0.2. 11. A crystal of lobaplatin compound according to claim 10, characterized in that the melting temperature T _like the specified crystal of the compound of lobaplatin is 230 ± 5 ° C. 12. The method of obtaining the specified crystal of lobaplatin according to claim 10 or 11, where the method includes the following stages: adding anhydrous methanol or anhydrous ethanol to lobaplatin trihydrate to obtain a mixture, stirring the mixture at room temperature to dissolve the solids, removing insoluble substances from the mixture, slowly evaporating the mixture, precipitating the crystal, isolating the crystal, and then drying the crystal to obtain a white powder, which represents a crystalline form B of lobaplatin. 13. The method according to p. 12, where the ratio of the mass of the specified lobaplatin trihydrate to the volume of anhydrous methanol is 1 (g): 40-50 (ml); the ratio of the mass of the specified lobaplatin trihydrate to the volume of anhydrous ethanol is 1 (g): 80-90 (ml). 14. A method of obtaining the specified crystal of lobaplatin according to claim 10 or 11, where the method includes the following stage b): adding anhydrous methanol to lobaplatin dihydrate to obtain a mixture, stirring the mixture at room temperature to dissolve the solids, removing insoluble substances from the mixture, slowly evaporating the mixture, precipitating the crystal, isolating the crystal, and then drying the crystal to obtain a white powder, which is a crystalline form B lobaplatin. 15. The method according to p. 14, where the method of obtaining the specified lobaplatin dihydrate comprises the following stage a): adding a crystallization solvent from the suspension to lobaplatin trihydrate to form a suspension, mixing the mixture, precipitating the crystal, washing with ethyl ether after removing the solvent, and then vacuum drying the crystal to obtain a crystal of lobaplatin dihydrate. 16. The method of claim 15, wherein said crystallization solvent in step a) is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane and wherein the weight ratio of said lobaplatin trihydrate to the volume of solvent for crystallization is 1 (g): 15-30 (ml). 17. The method according to any one of paragraphs. 14-16, where in step b), after isolation of the crystal, the crystal is washed with ethyl ether before drying the crystal, wherein said drying is vacuum drying. 18. The method according to any one of paragraphs. 14-16, where at the indicated stage b) the ratio of the mass of lobaplatin dihydrate to the volume of anhydrous methanol is 1 (g): 40-50 (ml). 19. The method of obtaining the specified crystal of lobaplatin according to claim 10 or 11, where the method includes the following stage b): adding an organic solvent to lobaplatin dihydrate to form a mixture in suspension, stirring the mixture at room temperature, precipitating a crystal, isolating a crystal, and then drying the crystal to obtain a white powder, which is crystalline form B of lobaplatin. 20. The method according to p. 19, where the method of obtaining the specified lobaplatin dihydrate comprises the following stage a): adding a solvent for crystallization from the suspension to lobaplatin trihydrate to form a suspension, mixing the mixture, precipitating the crystal and washing the crystal with ethyl ether after removing the solvent, and then vacuum drying the crystal to obtain a crystal of lobaplatin dihydrate. 21. The method of claim 20, wherein said crystallization solvent in step a) is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane, and wherein the weight ratio of said trihydrate lobaplatin to the volume of the crystallization solvent is 1 (g): 15-30 (ml). 22. The method according to any one of paragraphs. 19-21, where in step b), after separation of the crystal, the crystal is washed with ethyl ether before drying the crystal, wherein said drying is vacuum drying. 23. The method according to any one of paragraphs. 19-21, where the specified suspension in stage b) is obtained at room temperature, preferably the specified suspension is obtained within 45-50 hours 24. The method according to any one of paragraphs. 19-21, where the organic solvent in step b) is selected from n-hexane, acetone, ethyl acetate, nitromethane, acetonitrile, tetrahydrofuran, 2-butanone or dichloromethane and wherein the ratio of the mass of lobaplatin dihydrate to the volume of the organic solvent is 1 (g): 15-30 (ml). 25. Lobaplatin compound crystal, characterized in that the crystalline form of the compound crystal is crystalline form F and diffraction peaks are present on the PXRD pattern at 2θ values of about 8.21, 11.60, 12.99, 15.24, 16, 44, 17.11, 17.55, 18.42, 19.01, 19.20, 19.42, 21.81, 22.17, 22.42, 23.33, 23.85, 24.18, 24.40, 24.77, 25.46, 25.98, 26.13, 27.89, 28.42, 29.03, 30.32, 31.17, 31.94, 33.30, 36, 20, 37.62, 39.66, while the margin of error of the values of 2θ is 0.2. 26. The lobaplatin crystal according to claim 25, characterized in that the melting temperature T _etc. said crystal of lobaplatin compound is 229 ± 5 ° C. 27. A method of producing said lobaplatin crystal according to claim 25 or 26, characterized in that the method comprises the following step b): adding methanol or ethanol to lobaplatin dihydrate to obtain a mixture, stirring the mixture at room temperature to dissolve the solids, filtering the mixture to remove insoluble substances, slowly adding the organic solvent to the mixture, precipitating the crystal, isolating the crystal, and then drying the crystal to obtain a white powder, which is a crystalline form F of lobaplatin. 28. The method according to p. 27, where the method of obtaining the specified lobaplatin dihydrate comprises the following stage a): adding a suspension crystallization solvent to lobaplatin trihydrate to form a suspension, mixing the mixture, precipitating the crystal from the mixture, washing the crystal with ethyl ether after removing the solvent, and then vacuum drying the crystal to obtain a crystal of lobaplatin dihydrate. 29. The method of claim 28, wherein said crystallization solvent in step a) is selected from methyl tert-butyl ether, toluene, ethyl ether, butyl acetate, 1,4-dioxane or n-heptane and a weight ratio of said lobaplatin trihydrate to volume the crystallization solvent is 1 (g): 15-30 (ml). 30. The method according to any one of paragraphs. 27-29, where in step b), after isolation of the crystal, the crystal is washed with ethyl ether before drying the crystal, and wherein said drying is vacuum drying. 31. The method according to any one of paragraphs. 27-29, wherein said organic solvent in step b) is selected from dimethyl ether of ethylene glycol, n-hexane, ethyl acetate, acetone, nitromethane, acetonitrile, tetrahydrofuran or methylene chloride. 32. The method according to any one of paragraphs. 27-29, where the mass ratio of the specified lobaplatin dihydrate to the volume of the organic solvent in the indicated step b) is 1 (g): 120-200 (ml). 33. The method according to any one of paragraphs. 27-29, where at the indicated stage b) the mass ratio of the specified lobaplatin dihydrate to the volume of methanol is 1 (g): 40-50 (ml), the mass ratio of the specified lobaplatin dihydrate to the volume of ethanol is 1 (g): 80-90 (ml ) 34. A pharmaceutical composition, characterized in that said lobaplatin crystal according to any one of paragraphs. 1-4, 10, 11, 25 or 26 is used as the active ingredient. 35. The pharmaceutical composition according to p. 34, characterized in that the amount of lobaplatin crystal in the minimum unit of said pharmaceutical composition is 5, 10 or 50 mg. 36. The pharmaceutical composition according to p. 34 or 35, characterized in that the pharmaceutical composition is in any clinically acceptable pharmaceutical dosage form. 37. The pharmaceutical composition according to any one of paragraphs. 34, 35, characterized in that the dosage form is a lyophilized preparation for injection. 38. The use of the specified crystal lobaplatin according to any one of paragraphs. 1-4, 10, 11, 25 or 26 to obtain anti-cancer drugs. 39. The use of the specified pharmaceutical composition according to any one of paragraphs. 34-37 for obtaining anti-cancer drugs. 40. The use of the specified crystal lobaplatin according to any one of paragraphs. 1-4, 10, 11, 25, or 26 for the treatment of breast cancer, small cell lung cancer, or chronic myeloid leukemia. 41. The use of the specified pharmaceutical composition according to any one of paragraphs. 34-37 for the treatment of breast cancer, small cell lung cancer, or chronic myeloid leukemia.

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